Environmental DNA metabarcoding reflects spatiotemporal fish community shifts in the Scheldt estuary.

Estuarine ecosystems face increasing anthropogenic pressures, necessitating effective monitoring methods to mitigate their impacts on the biodiversity they harbour. The use of environmental DNA (eDNA) based detection methods is increasingly recognized as a promising tool to complement other, potentially invasive monitoring techniques. Integrating such eDNA analyses into monitoring frameworks for large ecosystems is still challenging and requires a deeper understanding of the scale and resolution at which eDNA patterns may offer insights in species presence and community composition space and time. The Scheldt estuary, characterized by its diverse habitats and complex currents, is one of the largest Western European tidal river systems. Until now, it remains challenging to obtain accurate information on fish communities living in and migrating through this ecosystem, consequently confining our knowledge to specific locations. To explore the potential of eDNA based monitoring, we simultaneously combine stow net fishing with eDNA metabarcoding, to assess spatiotemporal shifts in the Scheldt estuary's fish communities. In total, we detected 71 fish species in the estuary using eDNA metabarcoding, partly overlapping with historic fish community data gathered at the different study locations and in contrast to only 42 species using stow net fishing during the same survey period. Community compositions found by both detection methods varied among sampling locations, driven by a clear correlation to the salinity gradient. Limited effects of sampling depth and tide were observed on the eDNA metabarcoding data, allowing a significant reduction of the eDNA sampling effort for future eDNA fish monitoring campaigns in this study system. Our results further demonstrate that seasonal shifts in fish species occurrence can be detected using eDNA metabarcoding. Combining eDNA metabarcoding and stow net fishing further enhances our understanding of this vital waterway's diverse fish populations, allowing a higher resolution and more efficient monitoring strategy.

Using environmental DNA metabarcoding to monitor fish communities in small rivers and large brooks: Insights on the spatial scale of information.

Monitoring fish communities is central to the evaluation of ecological health of rivers. Both presence/absence of fish species and their relative quantity in local fish assemblages are crucial parameters to measure. Fish communities in lotic systems are traditionally monitored via electrofishing, characterized by a known limited efficiency and high survey costs. Analysis of environmental DNA could serve as a non-destructive alternative for detection and quantification of lotic fish communities, but this approach still requires further insights in practical sampling schemes incorporating transport and dilution of the eDNA particles; optimization of predictive power and quality assurance of the molecular detection method. Via a controlled cage experiment, we aim to extend the knowledge on streamreach of eDNA in small rivers and large brooks, as laid out in the European Water Framework Directive's water typology. Using a high and low source biomass in two river transects of a species-poor river characterized by contrasting river discharge rates, we found strong and significant correlations between the eDNA relative species abundances and the relative biomass per species in the cage community. Despite a decreasing correlation over distance, the underlying community composition remained stable from 25 to 300 m, or up to 1 km downstream of the eDNA source, depending on the river discharge rate. Such decrease in similarity between relative source biomass and the corresponding eDNA-based community profile with increasing distance downstream from the source, might be attributed to variation in species-specific eDNA persistence. Our findings offer crucial insights on eDNA behaviour and characterization of riverine fish communities. We conclude that water sampled from a relatively small river offers an adequate eDNA snapshot of the total fish community in the 300-1000 m upstream transect. The potential application for other river systems is further discussed.

Glass eel (Anguilla anguilla L. 1758) feeding behaviour during upstream migration in an artificial waterway.

The transition from marine to fresh water is a challenging task for juvenile eels. This critical step in the early eels' life is preceded by a metamorphosis from the oceanic larval to the continental glass eel stage, requiring major energy-demanding morphological, physiological and behavioural modifications during which time these animals do not feed. The success of the glass eels' inland migration after metamorphosis will largely depend on remaining energy levels, which can be supplemented only by resuming food uptake. Although it is crucial for their survival and the maintenance of the population, the feeding behaviour of glass eels is still an understudied aspect of the eels' complex life cycle. Many uncertainties about the phenology, diet, potential prey preferences and their relation with migration modus (migratory vs. sedentary) still remain. In this study, the authors analysed the stomach and gut contents of 458 European glass eels (Anguilla anguilla L. 1758) captured in a drainage canal connecting a small mesotidal estuary with an adjacent polder area during the spring migration seasons of 2016 and 2017. They demonstrated that although glass eels started feeding briefly upon arrival in the estuary, food uptake for early arrivals was restricted to a minority that sparsely feed on detritus and some worm-like benthic invertebrates. Along the season, food uptake intensified eventually engaging all glass eels and their dietary palette diversified including a wide array of planktonic and benthic organisms. Crustacean plankton (mainly cyclopoid copepods) was an important part of the glass eel diet, whereas benthic oligochaetes were less abundant as food source in spite of their high presence in the sediments. No clear differences in feeding behaviour could be observed between migratory and sedentary glass eels. This study showed that glass eels can use highly artificial and dynamic drainage canals as feeding ground during their critical marine/freshwater transition. This outcome is also a plea to improve the accessibility of alternative (unnatural) migration routes between the ocean and suitable freshwater growth habitats for the European eel.